Cell Surface Signalling Laboratory, Wellcome Sanger Institute, Cambridge, CB10 1SA, UK.
Stem Cell Genetics Laboratory, Wellcome Sanger Institute, Cambridge, CB10 1SA, UK.
Genome Biol. 2018 Nov 26;19(1):205. doi: 10.1186/s13059-018-1581-3.
Extracellular interactions between cell surface receptors are necessary for signaling and adhesion but identifying them remains technically challenging. We describe a cell-based genome-wide approach employing CRISPR activation to identify receptors for a defined ligand. We show receptors for high-affinity antibodies and low-affinity ligands can be unambiguously identified when used in pools or as individual binding probes. We apply this technique to identify ligands for the adhesion G-protein-coupled receptors and show that the Nogo myelin-associated inhibitory proteins are ligands for ADGRB1. This method will enable extracellular receptor-ligand identification on a genome-wide scale.
细胞表面受体之间的细胞外相互作用对于信号转导和黏附是必需的，但识别它们在技术上仍然具有挑战性。我们描述了一种基于细胞的全基因组方法，该方法利用 CRISPR 激活来鉴定针对特定配体的受体。我们表明，当高亲和力抗体和低亲和力配体用于池或作为单个结合探针时，可以明确鉴定出它们的受体。我们将此技术应用于鉴定黏附 G 蛋白偶联受体的配体，并表明 Nogo 髓鞘相关抑制蛋白是 ADGRB1 的配体。该方法将能够在全基因组范围内识别细胞外受体-配体。
